AI-Driven Virtual Screening and First-in-Class Conformational Biosensor Enable the Discovery of a GPR183 Inverse Agonist

ABSTRACT GPR183 is a chemotactic G protein-coupled receptor implicated in immune cell migration and various diseases. Here, we report the discovery of novel GPR183 inverse agonists using an artificial intelligence-driven virtual screening approach combined with biophysical sensors including a first-in-class conformational biosensor. From an initial screen of 70 compounds and a hit expansion, a selected candidate - compound 78 - demonstrated potent inhibition of GPR183’s constitutive and agonist-stimulated Gi activation as well β-arrestin2 recruitment. The binding of the compound to the receptor core was confirmed by studies using a new conformational sensor based on intramolecular BRET, molecular dynamics, and site-directed mutagenesis experiments. Finally, the compound 78 was able to block agonist-induced migration of peripheral blood mononuclear cells ex vivo . This work provides a novel chemical scaffold for GPR183 modulation. It introduces a new tool for dissecting receptor function, paving the way for therapeutic exploration in inflammatory, autoimmune, and neoplastic disorders.